Pharmaceutical formulation comprising an aqueous suspension of an androstane derivative for the treatment of inflammatory and allergic conditions

ABSTRACT

There is provided a pharmaceutical formulation comprising an aqueous suspension of particulate compound of formula (I)  
                 
or a solvate thereof.

This application is a Continuation-in-part of U.S. patent applicationSer. No. 09/958050 filed on 2 Oct. 2001, which is a 35 USC 371 USNational Phase of International Patent Application No. PCT.GB01.03495filed 3 Aug. 2001, which claims priority from United Kingdom PatentApplication No. GB 0019172.6 filed 5 Aug. 2000.

The present invention relates to pharmaceutical formulations containingan anti-inflammatory and anti-allergic compound of the androstane seriesand to processes for their preparation. The present invention alsorelates to therapeutic uses thereof, particularly for the treatment ofinflammatory and allergic conditions.

Glucocorticoids which have anti-inflammatory properties are known andare widely used for the treatment of inflammatory disorders or diseasessuch as asthma and rhinitis. For example, U.S. Pat. No. 4,335,121discloses6α,9α-Difluoro-17α-(1-oxopropoxy)-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester (known by the generic name of fluticasonepropionate) and derivatives thereof. The use of glucocorticoidsgenerally, and especially in children, has been limited in some quartersby concerns over potential side effects. The side effects that arefeared with glucocorticoids include suppression of theHypothalamic-Pituitary-Adrenal (HPA) axis, effects on bone growth inchildren and on bone density in the elderly, ocular complications(cataract formation and glaucoma) and skin atrophy. Certainglucocorticoid compounds also have complex paths of metabolism whereinthe production of active metabolites may make the pharmacodynamics andpharmacokinetics of such compounds difficult to understand. Whilst themodem glucocorticoids are very much safer than those originallyintroduced, it remains an object of research to produce new moleculesand formulations of old and new molecules which have excellentanti-inflammatory properties, with predictable pharmacokinetic andpharmacodynamic properties, with an attractive side effect profile, andwith a convenient treatment regime.

In PCT.GB01.03495 we have a novel glucocorticoid compound whichsubstantially meets these objectives. We have now identified aformulation of the novel glucocortoid compound in particular onesuitable for intranasal administration.

Many millions of individuals suffer from seasonal and perennial allergicrhinitis worldwide. Symptoms of seasonal and perennial allergic rhinitisinclude nasal itch, congestion, runny nose, sneezing and watery eyes.Seasonal allergic rhinitis is commonly known as ‘hay fever’. It iscaused by allergens which are present in the air at specific times ofthe year, for example tree pollen during Spring and Summer. Perennialallergic rhinitis is caused by allergens which are present in theenvironment during the entire year, for example dust mites, mould,mildew and pet dander.

To formulate an effective pharmaceutical nasal composition, themedicament must be delivered readily to all portions of the nasalcavities (the target tissues) where it performs its pharmacologicalfunction. Additionally, the medicament should remain in contact with thetarget tissues for relatively long periods of time. The longer themedicament remains in contact with the target tissues, the medicamentmust be capable of resisting those forces in the nasal passages thatfunction to remove particles from the nose. Such forces, referred to as‘mucociliary clearance’, are recognised as being extremely effective inremoving particles from the nose in a rapid manner, for example, within10-30 minutes from the time the particles enter the nose.

Other desired characteristics of a nasal composition are that it mustnot contain ingredients which cause the user discomfort, that it hassatisfactory stability and shelf-life properties, and that it does notinclude constituents that are considered to be detrimental to theenvironment, for example ozone depletors. In the case of administrationof glucocorticoids, the potential for any undesirable side effectsshould preferably be minimised.

Thus, according to one aspect of the invention, there is provided apharmaceutical formulation comprising an aqueous suspension ofparticulate compound of formula (I)

or a solvate thereof.

Preferably, the formulation will contain one or more suspending agents.

Preferably, the formulation will contain one or more preservatives.

Preferably, the formulation will contain one or more wetting agents.

Preferably, the formulation will contain one or more isotonicityadjusting agents.

According to one particular aspect of the present invention we provide apharmaceutical formulation which comprises:

-   -   (i) an aqueous suspension of particulate compound of formula (I)        or a solvate thereof;    -   (ii) one or more suspending agents;    -   (iii) one or more preservatives;    -   (iv) one or more wetting agents; and    -   (v) one or more isotonicity adjusting agents.

The formulations of the present invention may be stabilised byappropriate selection of pH. Typically, the pH will be adjusted tobetween 4.5 and 7.5, preferably between 5.0 and 7.0, especially around6.0.

Examples of pharmaceutically acceptable materials which can be used toadjust the pH of the formulation include hydrochloric acid and sodiumhydroxide. Preferably, the pH of the formulation will be adjusted usinghydrochloric acid.

The aqueous component is preferably a high grade quality of water, mostpreferably purified water.

The active compound of formula (I) or solvate thereofwill suitably havea mass mean diameter (MMD) of less than 20 μm, preferably between 0.5-10μm, especially between 1-5 μm. If particle size reduction is necessary,this may be achieved by techniques such as micronisation and/ormicrofluidisation.

Particularly preferred MMDs are between 2-4 μm.

In some preferred embodiments, if necessary, particle size reduction maybe achieved by micronisation.

In other preferred embodiments, particle size reduction may be achievedby microfluidisation.

Preferably, the particles will be crystalline, prepared for example by aprocess which comprises mixing in a continuous flow cell in the presenceof ultrasonic radiation a flowing solution of compound of formula (I) orsolvate thereof as medicament in a liquid solvent with a flowing liquidantisolvent for said medicament (as described in International PatentApplication PCT.GB99.04368).

A pharmaceutically effective amount of particulate compound of formula(I) or solvate thereof is present within the formulation, in an amountwhich is preferably between 0.005% and 1% (w/w), preferably between0.01% and 0.5% (w/w), especially 0.05-0.1% (w/w) based on the totalweight of the formulation. Typically, 50μl of suspension will contain50μg of compound of formula (I) or solvate thereof.

Examples of suspending agents include carboxymethylcellulose, veegum,tragacanth, bentonite, methylcellulose and polyethylene glycols.Preferably, the suspending agent will be microcrystalline cellulose andcarboxy methylcellulose sodium, most preferably used as the brandedproduct Avicel RC591 (which typically contains 87-91% microcrystallinecellulose and 9-13% carboxy methylcellulose sodium). Particulatemicrocrystalline cellulose will preferably have a particle size in therange 1 to 100 μm. We believe that Avicel RC591 acts as a suspendingagent by imparting thixotropic properties to the formulation, whereinthe formulation may become a stable suspension upon being stirred,shaken or otherwise disturbed.

Preferably, the thixotropic nature of the suspending agent will ensurethat the formulation assumes a gel like appearance at rest, wherein theparticulate medicament is dispersed and suspended substantiallyuniformly, characterised by a high viscosity value. Once the compositionis subjected to shear forces, such as those caused by agitation prior tospraying, the viscosity of the formulation will preferably decrease tosuch a level to enable it to flow readily through the spray device andexit as a spray of fine particles in a mist. These particles will thenbe capable of infiltrating the mucosal surfaces of the anterior regionsof the nose (frontal nasal cavities), the frontal sinus, the maxillarysinuses and the turbinates which overlie the conchas of the nasalcavities. Once deposited, the viscosity of the formulation willpreferably increase to a sufficient level to assume its gel-like formand resist being cleared from the nasal passages by the inherentmucocillary forces that are present in the nasal cavities.

When the formulation of the present invention comprises a suspendingagent, it will be desirably added in a suitable amount to achieve thisfunction, preferably the suspending agent will be present within theformulation in an amount of between 0.1 and 5% (w/w), especially 1.5%(w/w), based on the total weight of the formulation.

For stability purposes, the formulation of the present invention shouldbe protected from microbial contamination and growth by inclusion of apreservative. Examples of pharmaceutically acceptable anti-microbialagents or preservatives that can be used in the formulation includequaternary ammonium compounds (eg. benzalkonium chloride, benzethoniumchloride, cetrimide and cetylpyridinium chloride), mercurial agents (eg.phenylmercuric nitrate, phenylmercuric acetate and thimerosal),alcoholic agents (eg. chlorobutanol, phenylethyl alcohol and benzylalcohol), antibacterial esters (eg. esters of para-hydroxybenzoic acid),chelating agents such as disodium edetate (EDTA) and otheranti-microbial agents such as chlorhexidine, chlorocresol, sorbic acidand its salts and polymyxin.

Preferably the preservative will comprise disodium edetate (EDTA), whichwill preferably be present within the formulation in an amount ofbetween 0.001 and 1% (w/w), especially around 0.015% (w/w), based on thetotal weight of the formulation.

Preferably, the preservative will comprise benzalkonium chloride (BKC),which will preferably be present within the formulation in an amount ofbetween 0.001 and 1% (w/w), especially around 0.015% (w/w), based on thetotal weight of the formulation.

More preferably, the preservative comprises disodium edetate andbenzalkonium chloride or disodium edetate and potassium sorbate,particularly disodium edetate and benzalkonium chloride.

Formulations, eg nasal formulations which contain a suspended medicament(such as a compound of formula (I) or a solvate thereof) will preferablycontain a pharmaceutically acceptable wetting agent which functions towet the particles of medicament to facilitate dispersion thereof in theaqueous phase of the composition. Preferably, the amount of wettingagent used will not cause foaming of the dispersion during mixing.

It will be appreciated that any agent which is effective in wetting theparticles and which is pharmaceutically acceptable can be used. Examplesof wetting agents that can be used are fatty alcohols, esters andethers. Preferably, the wetting agent will be a hydrophilic, non-ionicsurfactant, most preferably polyoxyethylene (20) sorbitan monooleate(supplied as the branded product Polysorbate 80).

Wherein the formulation of the present invention comprises a wettingagent, it will be desirably added in a sufficient quantity to achievethis function, preferably the wetting agent will be present within theformulation in an amount of between 0.001 and 0.05% (w/w), especially0.025% (w/w), based on the total weight of the formulation.

The presence of an isotonicity adjusting agent is to achieve isotonicitywith body fluids eg fluids of the nasal cavity, resulting in reducedlevels of irritancy associated with many nasal formulations. Examples ofsuitable isotonicity adjusting agents are sodium chloride, dextrose andcalcium chloride. Preferably, the isotonicity adjusting agent will bedextrose, most preferably used as anhydrous dextrose.

When the formulation of the present invention comprises an isotonicityadjusting agent it will be desirably added in a sufficient quantity toachieve this function, preferably the isotonicity adjusting agent willbe present within the formulation in an amount of between 0.1 and 10%(w/w), especially 5.0% w/w, based on the total weight of theformulation.

In the pharmaceutical formulation of the invention, the compound offormula (I) or solvate thereof is present within the formulation in anamount between 0.005% and 1% (w/w), based on the total weight of theformulation.

In the pharmaceutical formulation of the invention, preferably thesuspending agent is microcrystalline cellulose and carboxymethylcellulose sodium, the preservative is EDTA and benzalkoniumchloride, the wetting agent is polyoxyethylene (20) sorbitan monooleateand the isotonicity adjusting agent is dextrose.

The compound of formula (I) or a solvate thereof and formulationsthereof have potentially beneficial anti-inflammatory or anti-allergiceffects, particularly upon topical administration to the nose,demonstrated by, for example, its ability to bind to the glucocorticoidreceptor and to illicit a response via that receptor, with long actingeffect. Hence, formulations according to the invention are useful in thetreatment of inflammatory and/or allergic disorders of the nose,especially in once-per-day therapy.

Formulations according to the invention may be prepared by combining theingredients in water. If necessary the pH may be adjusted as a finalstep. Formulations so prepared may then be filled into the receptacle.

Aqueous formulations of the invention may also be employed for rectal,aural, otic, oral, topical or parenteral administration oradministration by inhalation for the treatment of other localinflammatory conditions (eg dermatitis, asthma, chronic obstructivepulmonary disease (COPD) and the like). For example formulations of theinvention may be administered to the lung by nebulisation. Suchformulations may employ excipients (eg preservatives, buffers and thelike) appropriate for the route of administration.

The particularly desirable biological properties of the compound offormula (I) are now explained below:

Compound or formula (I) undergoes highly efficient hepatic metabolism toyield the 17-β carboxylic acid (X) as the sole major metabolite in (atand human in vitro systems. This metabolite has been synthesised anddemonstrated to be >1000 fold less active than the parent compound in invitro functional glucocortcoid assays.

This efficient hepatic metabolism is reflected by in vivo data in therat, which have demonstrated plasma clearance at a rate approachinghepatic blood flow and an oral bioavailability of <1%, consistent withextensive first-pass metabolism.

In vitro metabolism studies in human hepatocytes have demonstrated thatcompound (I) is metabolised in a similar manner to fluticasonepropionate but that conversion of (I) to the inactive acid metaboliteoccurs approximately 5-fold more rapidly than with fluticasonepropionate. This very efficient hepatic inactivation would be expectedto minimise systemic exposure in man leading to an improved safetyprofile.

Examples of disease states in which the compound of formula (I) hasutility include inflammatory and/or allergic conditions of the nasalpassages such as rhinitis eg seasonal and perennial rhinitis as well asother local inflammatory conditions such as asthma, COPD and dermatitis.

It will be appreciated by those skilled in the art that reference hereinto treatment extends to prophylaxis as well as the treatment ofestablished conditions.

Preferable means for applying the formulation of the present inventionto the nasal passages is by use of a pre-compression pump, such as aVP3, VP7 or modifications, model manufactured by Valois SA. Advantagesof pumps of this type are beneficial as they will ensure that theformulation is not released or atomised until a sufficient force hasbeen applied, otherwise smaller doses may be applied. Typically, thesepre-compression pumps may be used with a bottle (glass or plastic)capable of holding 8-50 ml of a formulation. Each spray will typicallydeliver 50-100 μl of such a formulation, therefore, the device iscapable of providing at least 100 metered doses.

A suitable dosing regime for the formulation of the present inventionwhen administered to the nose would be for the patient to inhale slowlythrough the nose subsequent to the nasal cavity being cleared. Duringinhalation the formulation would be applied to one nostril while theother is manually compressed. This procedure would then be repeated forthe other nostril.

Typically, one or two inhalations per nostril would be administered bythe above procedure up to three times each day, ideally once daily.

It will be appreciated that the above dosing regime should be adjustedaccording to the patient's age, body weight and/or symptom severity.

As mentioned above, formulations comprising a compound of formula (I) orsolvate thereof are useful in human or veterinary medicine, inparticular as an anti-inflammatory and anti-allergic agent.

There is thus provided as a further aspect of the invention aformulation comprising the compound of formula (I) or solvate thereoffor use in human or veterinary medicine, particularly in the treatmentof patients with inflammatory and/or allergic conditions.

According to another aspect of the invention, there is provided the useof a formulation comprising the compound of formula (I) or solvatethereof for the manufacture of a medicament for the treatment ofpatients with inflammatory and/or allergic conditions.

In a further or alternative aspect, there is provided a method for thetreatment of a human or animal subject with an inflammatory and/orallergic condition, which method comprises administering to said humanor animal subject an effective amount of a formulation comprising thecompound of formula (I) or solvate thereof.

The compound of formula (I) is long-acting, therefore preferably thecompound will be delivered once-per-day and the dose will be selected sothat the compound has a therapeutic effect in the treatment ofrespiratory disorders (eg rhinitis) over 24 hours or more.

The pharmaceutical compositions according to the invention may also beused in combination with another therapeutically active agent, forexample, an anti-histamine or an anti-allergic. The invention thusprovides, in a further aspect, a combination comprising a compound offormula (I) or a physiologically acceptable salt or solvate thereoftogether with another therapeutically active agent, for example, ananti-histamine or an anti-allergic.

Examples of anti-histamines include methapyrilene or loratadine.

Other suitable combinations include, for example, otheranti-inflammatory agents eg. NSAIDs (eg. PDE4 inhibitors, leukotrieneantagonists, iNOS inhibitors, tryptase and elastase inhibitors, beta-2integrin antagonists and adenosine 2a agonists)) or antiinfective agents(eg. antibiotics, antivirals).

Of particular interest is use of the compounds of formula (I) incombination with a phosphodiesterase 4 (PDE4) inhibitor. ThePDE4-specific inhibitor useful in this aspect of the invention may beany compound that is known to inhibit the PDE4 enzyme or which isdiscovered to act as a PDE4 inhibitor, and which are only PDE4inhibitors, not compounds which inhibit other members of the PDE familyas well as PDE4. Generally it is preferred to use a PDE4 inhibitor whichhas an IC₅₀ ratio of about 0.1 or greater as regards the IC₅₀ for thePDE4 catalytic form which binds rolipram with a high affinity divided bythe IC₅₀ for the form which binds rolipram with a low affinity. For thepurposes of this disclosure, the CAMP catalytic site which binds R and Srolipram with a low affinity is denominated the “low affinity” bindingsite (LPDE 4) and the other form of this catalytic site which bindsrolipram with a high affinity is denominated the “high affinity” bindingsite (HPDE 4). This term “HPDE4” should not be confused with the term“hPDE4” which is used to denote human PDE4.

The pharmaceutical formulation according to the invention may furthercomprise one or more excipients. By the term “excipient”, as usedherein, it is meant to mean substantially inert materials that arenontoxic and do not interact with other components of a composition in adeleterious manner including, but not limited to, pharmaceutical gradesof: carbohydrates, organic and inorganic salts, polymers, amino acids,phospholipids, wetting agents, emulsifiers, surfactants, poloxamers,pluronics, and ion exchange resins, and combinations thereof, anon-exhaustive list of examples of which are provided below:

Carbohydrates, including: monosaccharides, such as, but not limited to,fructose; disaccharides, such as, but not limited to lactose, andcombinations and derivatives thereof; polysaccharides, such as, but notlimited to, cellulose and combinations and derivatives thereof;oligosaccharides, such as, but not limited to, dextrins, andcombinations and derivatives thereof; polyols, such as but not limitedto sorbitol, and combinations and derivatives thereof;

Organic and inorganic salts, including but not limited to sodium orcalcium phosphates, magnesium stearate, and combinations and derivativesthereof;

Polymers, including: natural biodegradable protein polymers including,but not limited to, gelatin and combinations and derivatives thereof;

Natural biodegradable polysaccharide polymers including, but not limitedto, chitin and starch, crosslinked starch, and combinations andderivatives thereof;

Semisynthetic biodegradable polymers including, but not limited to,derivatives of chitosan;

Synthetic biodegradable polymers including but not limited topolyethylene glycols (PEG), polylactic acid (PLA), synthetic polymersincluding but not limited to polyvinyl alcohol and combinations andderivatives thereof;

-   -   Amino acids including but not limited to including non-polar        amino acids, such as leucine and combinations and derivatives        thereof;    -   Phospholipids, including lecithins and combinations and        derivatives thereof;    -   Wetting agents/Surfactants/Emulsifiers, including, but not        limited to gum acacia, cholesterol, fatty acids including,        combinations and derivatives thereof;    -   Poloxamers/Pluronics: including but not limited to poloxamer        188, Pluronic® F-108, and combinations and derivations thereof;    -   Ion exchange resins: including but not limited to amberlite        IR120 and combinations and derivatives thereof,    -   and combinations of the noted excipients.

Further, there is provided a process for the preparation of suchpharmaceutical compositions which comprises mixing the ingredients.

The individual compounds of such combinations may be administered eithersequentially in separate pharmaceutical compositions as well assimultaneously in combined pharmaceutical formulations. Preferablyadditional therapeutically active ingredients are suspended in theformulation together with the compound of formula (I). Appropriate dosesof known therapeutic agents will be readily appreciated by those skilledin the art

A process for preparing a compound of formula (I) comprises alkylationof a thioacid of formula (II)

or a salt thereof.

In this process the compound of formula (II) may be reacted with acompound of formula FCH₂L wherein L represents a leaving group (eg ahalogen atom, a mesyl or tosyl group or the like) for example, anappropriate fluoromethyl halide under standard conditions. Preferably,the fluoromethyl halide reagent is bromofluoromethane. Preferably thecompound of formula (II) is employed as a salt, particularly the saltwith diisopropylethylamine.

In a preferred process for preparing the compound of formula (I), thecompound of formula (II) or a salt thereof is treated withbromofluoromethane optionally in the presence of a phase transfercatalyst A preferred solvent is methylacetate, or more preferablyethylacetate, optionally in the presence of water. The presence of waterimproves solubility of both starting material and product and the use ofa phase transfer catalyst results in an increased rate of reaction.Examples of phase transfer catalysts that may be employed include (butare not restricted to) tetrabutylammonium bromide, tetrabutylammoniumchloride, benzyltributylammonium bromide, benzyltributylammoniumchloride, benzyltriethylammonium bromide, methyltributylammoniumchloride and methyltrioctylammonium chloride. THF has also successfullybeen employed as solvent for the reaction wherein the presence of aphase transfer catalyst again provides a significantly faster reactionrate. Preferably the product present in an organic phase is washedfirstly with aqueous acid eg dilute HCl in order to remove aminecompounds such as triethylamine and diisopropylethylamine and then withaqueous base eg sodium bicarbonate in order to remove any unreactedprecursor compound of formula (II).

Compound of formula (i) in unsolvated form may be prepared by a processcomprising:

-   -   (a) Crystallising the compound of formula (I) in the presence of        a non-solvating solvent such as ethanol, methanol, water, ethyl        acetate, toluene, methylisobutylketone or mixtures thereof; or    -   (b) Desolvating a compound of formula (I) in solvated form (eg        in the form of a solvate with acetone, isopropanol,        methylethylketone, DMF or tetrahydrofuran) eg by heating.

In step (b) the desolvation will generally be performed at a temperatureexceeding 50° C. preferably at a temperature exceeding 100° C. Generallyheating will be performed under vacuum.

Compound of formula (I) in unsolvated form has been found to exist in 3crystalline polymorphic forms, Forms 1, 2 and 3. The Forms arecharacterised by their X-ray diffraction (XRPD) patterns Broadlyspeaking the Forms are characterised. in their XRPD profiles as follows:

-   Form 1: Peak at around 18.9 degrees 2Theta-   Form 2: Peaks at around 18.4 and 21.5 degrees 2Theta-   Form 3: Peaks at around 18.6 and 19.2 degrees 2Theta.

Unsolvated polymorph Form 1 appears likely to be the thermodynamicallymost stable form since Forms 2 and 3 are converted into Form I onheating. Polymorph Form 1 is preferred.

In some embodiments we prefer the compound of formula (I) in unsolvatedpolymorph Form 1 substantially to be in the form of equant orsubstantially equant particles. Substantially equant particles havedimensions in the X, Y and Z dimensions which are similar in length.

Another aspect of the present invention provides compounds of formula(I) substantially in the form of equant or substantially equantparticles.

Preferably, the equant particles will be in the form of tetragonalbipyramidal or substantially tetragonal bipyramidal particles.

Tetragonal bipyramidal particles of Form 1 polymorph have surprisinglybeen found to produce relatively consistent sized particles aftermicronisation, which appears to be independent of the drying processprior to micronisation. The tetragonal bipyramidal particles possessgood flow properties, good bulk density, and are easily isolated fromsuspension by filtration.

In other embodiments we prefer the compound of formula (I) in unsolvatedpolymorph Form 1 to be in the form of acicular particles. Acicularparticles are needle-like in shape.

Another aspect of the present invention provides compounds of formula(I) substantially in the form of acicular particles.

Acicular particles of Form 1 have surprisingly been found to producerelatively varying sized particle distribution after micronisation, andparticle sizes which, on average, are larger than those produced frommicronisation of tetragonal bipyramidal particles. In certaincircumstances where this size profile of micronised product ispreferred, then acicular particles will be preferred. It may be that themicronised product produced from acicular particles will result in lowersystemic exposure to the active compound.

A process for preparing a compound of formula (I) as unsolvated Form 1polymorph comprises dissolving compound of formula (I) inmethylisobutylketone, ethyl acetate or methyl acetate and producingcompound of formula (I) as unsolvated Form 1 by addition of anon-solvating anti-solvent such as iso-octane or toluene.

According to a first preferred embodiment of this process the compoundof formula (I) may be dissolved in ethyl acetate and compound of formula(I) as unsolvated Form I polymorph may be obtained by addition oftoluene as anti-solvent In order to improve the yield, preferably theethyl acetate solution is hot and once the toluene has been added themixture is distilled to reduce the content of ethyl acetate.

According to a second preferred embodiment of this process the compoundof formula (I) may be dissolved in methylisobutylketone and compound offormula (I) as unsolvated Form 1 polymorph may be obtained by additionof isooctane as anti-solvent

Compound of formula (I) in solvated form may be prepared bycrystallising the compound of formula (I) from a solvating solvent suchas acetone, propan-2-ol, ethylacetate or tetrahydrofuran (THF).

Preferably in processes for preparing formulations of the invention, thecompound of formula (I) will be employed in unsolvated form, typicallyunsolvated Form 1.

Thus, according to another aspect of the present invention there isprovided a process for the preparation of a compound of formula (I),substantially in the form of equant or substantially equant particleswhich process comprises dissolving a compound of formula (I) in asuitable solvating solvent and crystallising the compound

Preferred solvating agents are acetone, propan-2-ol, ethylacetate ortetrahydrofuran, more preferably acetone or propan-2-ol.

The compound of formula (I) is preferably substantially in the form oftetragonal bipyramidel particles.

Compounds of formula (II) may be prepared from the corresponding17α-hydroxyl derivative of formula (III):

using for example, the methodology described by G. H. Phillipps et al.,(1994) Journal of Medicinal Chemistry, 37, 3717-3729. For example thestep typically comprises the addition of a reagent suitable forperforming the esterification eg an activated derivative of 2-furoicacid such as an activated ester or preferably a 2-furoyl halide eg2-furoyl chloride (employed in at least 2 times molar quantity relativeto the compound of formula (III)) in the presence of an organic base egtriethylamine. The second mole of 2-furoyl chloride reacts with thethioacid moiety in the compound of formula (III) and needs to be removedeg by reaction with an amine such as diethylamine.

This method suffers disadvantages, however, in that the resultantcompound of formula (II) is not readily purified of contamination withthe by-product 2-furoyldiethylamide. We have therefore invented severalimproved processes for performing this conversion.

In a first such improved process we have discovered that by using a morepolar amine such as diethanolamine, a more water soluble by-product isobtained (in this case 2-furoyldiethanolamide) which permits compound offormula (II) or a salt thereof to be produced in high purity since theby-product can efficiently be removed by water washing.

Thus we provide a process for preparing a compound of formula (II) whichcomprises:

-   -   (a) reacting a compound of formula (III) with an activated        derivative of 2-furoic acid as in an amount of at least 2 moles        of the activated derivative per mole of compound of        formula (III) to yield a compound of formula (IIA);        and    -   (b) removal of the sulphur-linked 2-furoyl moiety from compound        of formula (IIA) by reaction of the product of step (a) with an        organic primary or secondary amine base capable of forming a        water soluble 2-furoyl amide.

In two particularly convenient embodiments of this process we alsoprovide methods for the efficient purification of the end product whichcomprise either

-   -   (c1)when the product of step (b) is dissolved in a substantially        water immiscible organic solvent, purifying the compound of        formula (II) by washing out the amide by-product from step (b)        with an aqueous wash, or    -   (c2) when the product of step (b) is dissolved in a water        miscible solvent, purifying the compound of formula (II) by        treating the product of step (b) with an aqueous medium so as to        precipitate out pure compound of formula (II) or a salt thereof;

In step (a) preferably the activated derivative of 2-furoic acid may bean activated ester of 2-furoic acid, but is more preferably a 2-furoylhalide, especially 2-furoyl chloride. A suitable solvent for thisreaction is ethylacetate or methylacetate (preferably methylacetate)(when step (c1) may be followed) or acetone (when step (c2) may befollowed). Normally an organic base eg triethylamine will be present. Instep (b) preferably the organic base is diethanolamine. The base maysuitably be dissolved in a solvent eg methanol. Generally steps (a) and(b) will be performed at reduced temperature eg between 0 and 5° C. Instep (c1) the aqueous wash may be water, however the use of brineresults in higher yields and is therefore preferred. In step (c2) theaqueous medium is for example a dilute aqueous acid such as dilute HCl.

We also provide an alternative process for preparing a compound offormula (II) which comprises:

-   -   (a) reacting a compound of formula (III) with an activated        derivative of 2-furoic acid in an amount of at least 2 moles of        activated derivative per mole of compound of formula (III) to        yield a compound of formula (IIA); and    -   (b) removal of the sulphur-linked 2-furoyl moiety from compound        of formula (IIA) by reaction of the product of step (a) with a        further mole of compound of formula (III) to give two moles of        compound of formula (II).

In step (a) preferably the activated derivative of 2-furoic acid may bean activated ester of 2-furoic acid, but is more preferably a 2-furoylhalide, especially 2-furoyl chloride. A suitable solvent for his step isacetone. Normally an organic base eg triethylamine will be present. Instep (b) a suitable solvent is DMF or dimethylacetamide. Normally anorganic base eg triethylamine will be present. Generally steps (a) and(b) will be performed at reduced temperature eg between 0 and 5° C. Theproduct may be isolated by treatment with acid and washing with water.

This aforementioned process is very efficient in that it does notproduce any furoylamide by-product (thus affording inter aliaenvironmental advantages) since the excess mole of furoyl moiety istaken up by reaction with a further mole of compound of formula (II) toform an additional mole of compound of formula (II).

Further general conditions for the conversion of compound of formula(III) to compound of formula (II) in the two processes just describedwill be well known to persons skilled in the art.

According to a preferred set of conditions, however, we have found thatthe compound of formula (II) may advantageously be isolated in the formof a solid crystalline salt. The preferred salt is a salt formed with abase such as triethylamine, 2,4,6-trimethylpyridine,diisopropylethylamine or N-ethylpiperidine. Such salt forms of compoundof formula (II) are more stable, more readily filtered and dried and canbe isolated in higher purity than the free thioacid. The most preferredsalt is the salt formed with diisopropylethylamine. The triethylaminesalt is also of interest.

Compounds of formula (III) may be prepared in accordance with proceduresdescribed in GB 2088877B. Compounds of formula (III) may also beprepared by a process comprising the following steps:

Step (a) comprises oxidation of a solution containing the compound offormula (V). Preferably, step (a) will be performed in the presence of asolvent comprising methanol, water, tetrahydrofuran, dioxan ordiethylene glygol dimethylether. So as to enhance yield and throughput,preferred solvents are methanol, water or tetrahydrofuran, and morepreferably are water or tetrahydrofuran, especially water andtetrahydrofuran as solvent. Dioxan and diethylene glygol dimethyletherare also preferred solvents which may optionally (and preferably) beemployed together with water. Preferably, the solvent will be present inan amount of between 3 and 10 vol relative to the amount of the startingmaterial (1 wt.), more preferably between 4 and 6 vol., especially 5vol. Preferably the oxidising agent is present in an amount of 1-9 molarequivalents relative to the amount of the starting material. Forexample, when a 50% w/w aqueous solution of periodic acid is employed,the oxidising agent may be present in an amount of between 1.1 and 10wt. relative to the amount of the starting material (1 wt), morepreferably between 1.1 and 3 wt., especially 1.3 wt Preferably, theoxidation step will comprise the use of a chemical oxidising agent. Morepreferably, the oxidising agent will be periodic acid or iodic acid or asalt thereof. Most preferably, the oxidising agent will be periodic acidor sodium periodate, especially periodic acid. Alternatively (or inaddition), it will also be appreciated that the oxidation step maycomprise any suitable oxidation reaction, eg one which utilises airand/or oxygen. When the oxidation reaction utilises air and/or oxygen,the solvent used in said reaction will preferably be methanol.Preferably, step (a) will involve incubating the reagents at roomtemperature or a little warmer, say around 25° C. eg for 2 hours. Thecompound of formula (IV) may be isolated by recrystallisation from thereaction mixture by addition of an anti-solvent. A suitable anti-solventfor compound of formula (IV) is water. Surprisingly we have discoveredthat it is highly desirable to control the conditions under which thecompound of formula (IV) is precipitated by addition of anti-solvent egwater. When the recrystallisation is performed using chilled water (egwater/ice mixture at a temperature of 0-5° C.) although betteranti-solvent properties may be expected we have found that thecrystalline product produced is very voluminous, resembles a soft geland is very difficult to filter. Without being limited by theory webelieve that this low density product contains a large amount ofsolvated solvent within the crystal lattice. By contrast when conditionsof around 10° C. or higher are used (eg around ambient temperature) agranular product of a sand like consistency which is very easilyfiltered is produced. Under these conditions, crystallisation typicallycommences after around 1 hour and is typically completed within a fewhours (eg 2 hours). Without being limited by theory we believe that thisgranular product contains little or no solvated solvent within thecrystal lattice.

Step (b) will typically comprise the addition of a reagent suitable forconverting a carboxylic acid to a carbothioic acid eg using hydrogensulphide gas together with a suitable coupling agent egcarbonyldiimidazole (CDI) in the presence of a suitable solvent egdimethylformamide.

The advantages of the formulation of the compound of formula (I)according to the invention may include the fact that the formulationsdemonstrate excellent anti-inflammatory properties, with predictablepharmacokinetic and pharmacodynamic behaviour, with an attractiveside-effect profile, rapid onset of action, long duration of action, andare compatible with a convenient regime of treatment in human patients,in particular being amendable to once-per day dosing. Further advantagesmay include the fact that the formulation has desirable physical andchemical properties which allow for ready manufacture and storage.

According to another aspect of the invention there is provided acontainer comprising a pharmaceutical formulation according to theinvention.

There is also provided a device adapted for intranasal delivery of apharmaceutical formulation comprising such a container

Suitably the formulation will be dispensed from a vessel fitted with asuitable pre-compression pump and nasal actuator, adapted to dispense 50or 100 μl per actuation, preferably 50 μl

The following non-limiting Examples illustrate the invention:

EXAMPLES

General

¹H-nmr spectra were recorded at 400 MHz and the chemical shifts areexpressed in ppm relative to tetramethylsilane. The followingabbreviations are used to describe the multiplicities of the signals: s(singlet), d (doublet), t (triplet), q (quartet), m (multiplet), dd(doublet of doublets), ddd (doublet of doublet of doublets), dt (doubletof triplets) and b (broad). Biotage refers to prepacked silica gelcartridges containing KP-Sil run on flash 12 i chromatography module.LCMS was conducted on a Supelcosil LCABZ+PLUS column (3.3 cm×4.6 mm ID)eluting with 0.1% HCO₂H and 0.01 M ammonium acetate in water (solventA), and 0.05% HCO₂H 5% water in acetonitrile (solvent B), using thefollowing elution gradient 0-0.7 min 0%B, 0.7-4.2 min 100%B, 4.2-5.3 min0%B, 5.3-5.5 min 0%B at a flow rate of 3 ml/min. The mass spectra wererecorded on a Fisons VG Platform spectrometer using electrospraypositive and negative mode (ES+ve and ES−ve).

Intermediates

Intermediate 1:6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicAcid Diisopropylethylamine Salt

A stirred suspension of6α,9α-difluoro-11β,17α-dihydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid (prepared in accordance with the procedure described in GB2088877B) (49.5 g) in methylacetate (500 ml) is treated withtriethylamine (35 ml) maintaining a reaction temperature in the range0-5° C. 2-Furoyl chloride (25 ml) is added and the mixture stirred at0-5° C. for 1 hour. A solution of diethanolamine (52.8 g) in methanol(50 ml) is added and the mixture stirred at 0-5° C. for at least 2hours. Dilute hydrochloric acid (approx 1M, 550 ml) is added maintaininga reaction temperature below 15° C. and the mixture stirred at 15° C.The organic phase is separated and the aqueous phase is back extractedwith methyl acetate (2×250 ml). All of the organic phases are combined,washed sequentially with brine (5×250 ml) and treated withdi-isopropylethylamine (30 ml). The reaction mixture is concentrated bydistillation at atmospheric pressure to an approximate volume of 250 mland cooled to 25-30° C. (crystallisation of the desired product normallyoccurs during distillation/subsequent cooling). Tertiary butyl methylether (TBME) (500 ml) is added, the slurry further cooled and aged at0-5° C. for at least 10 minutes. The product is filtered off, washedwith chilled TBME (2×200 ml) and dried under vacuum at approximately40-50° C. (75.3 g, 98.7%). NMR (CDCl₃) δ: 7.54-7.46 (1H, m), 7.20-7.12(1H, dd), 7.07-6.99 (1H, dd), 6.48-6.41 (2H, m), 6.41-6.32 (1H, dd),5.51-5.28 (1H, dddd ²J_(H-F)50 Hz), 4.45-4.33(1H, bd), 3.92-3.73 (3H,bm), 3.27-3.14 (2H, q), 2.64-2.12 (5H, m), 1.88-1.71 (2H, m), 1.58-1.15(3H, s), 1.50-1.38 (15H, m), 1.32-1.23 (1H, m), 1.23-1.15 (3H s),1.09-0.99 (3H, d)

Intermediate 2:6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3oxo-androsta-1,4-diene-17β-carbothioicAcid S-fluoromethyl Ester Unsolvated Form 1

A mobile suspension of Intermediate 1 (12.61 g, 19.8 mmol) in ethylacetate (230 ml) and water (50 ml) is treated with a phase transfercatalyst (benzyltributylammonium chloride, 10 mol %), cooled to 3° C.and treated with bromofluoromethane (1.10 ml, 19.5 mmol, 0.98equivalents), washing in with prechilled (0° C.) ethyl acetate (EtOAc)(20 ml). The suspension is stirred overnight, allowing to warm to 17° C.The aqueous layer is separated and the organic phase is sequentiallywashed with 1M HCl (50 ml), 1% w/v NaHCO₃ solution (3×50 ml) and water(2×50 ml). The ethylacetate solution is distilled at atmosphericpressure until the distillate reaches a temperature of approximately 73°C. at which point toluene (150 ml) is added. Distillation is continuedat atmospheric pressure until all remaining EtOAc has been removed(approximate distillate temperature 103° C.). The resultant suspensionis cooled and aged at <10° C. and filtered off. The bed is washed withtoluene (2×30 ml) and the product oven dried under vacuum at 60° C. toconstant weight to yield the title compound (8.77 g, 82%) LCMS retentiontime 3.66 min, m/z 539 MH⁺, NMR δ (CDCl₃) includes 7.60 (1H, m),7.18-7.11 (2H, m), 6.52 (1H, dd, J 4.2 Hz), 6.46 (1H, s), 6.41 (1H, dd,J 10, 2 Hz), 5.95 and 5.82 (2H dd, J 51, 9 Hz), 5.48 and 5.35 (1H, 2m),4.48 (1H, m), 3.48 (1H, m), 1.55 (3H, s), 1.16 (3H, s), 1.06 (3H, d, J 7Hz).

Intermediate 3:6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicAcid S-fluoromethyl Ester Unsolvated Form 1 in Tetragonal BipyramidalHabit

A sample of Intermediate 2 (1 wt) is suspended in a mixture ofpropan-2-ol (2 vols), water (1 vol), ethyl acetate (13 vols). Thesuspension is heated to reflux to give a solution. The solution isclarified by passing a 5-micron line filter, following with a line washof ethyl acetate (2 vols). The solution is concentrated to ca 10 volsvia distillation at atmospheric pressure. Propan-2-ol (10 vols) isadded. The solution is seeded with the propan-2-ol solvate of6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester (0.01 wt) suspended in propan-2-ol (ca 0.03vols). The suspension is adjusted to 50+5° and aged for ca 5 mins (untilcrystallisation is established). The suspension is concentrated onceagain to ca 10 vols. Further propan-2-ol (10 vols) is added and thesuspension concentrated to ca 10 vols for a third time (note: pot tempat this point ca80°). The suspension is cooled to 0-5° C. over ca 120mins and aged for at least 60 mins. The white precipitate is isolated byvacuum filtration washing the filter cake with chilled propan-2-ol anddried in vacuo to afford the propan-2-ol solvate as an off-white to greysolid.

Stoichiometry of compound of formula (I): propan-2-ol=1:1 from ¹H nmr(CDCl₃)

The propan-2-ol is removed by the following process:

The product is heated under vacuum at 105 to 115° C. for at least 12hours to give tetragonal bipyramidal particles of unsolvated Form 1 ofthe active compound.

Pharmacological A

In Vitro Pharmacological Activity

Pharmacological activity was assessed in a functional in vitro assay ofglucocorticoid agonist activity which is generally predictive ofanti-inflammatory or anti-allergic activity in vivo.

For the experiments in this section, compound of formula (I) was used asunsolvated Form 1 (Intermediate 2)

The functional assay was based on that described by K. P. Ray et al.,Biochem J. (1997), 328, 707-715. A549 cells stably transfected with areporter gene containing the NF-κB responsive elements from the ELAMgene promoter coupled to sPAP (secreted alkaline phosphatase) weretreated with test compounds at appropriate doses for 1 hour at 37° C.The cells were then stimulated with tumour necrosis factor (TNF, 10ng/ml) for 16 hours, at which time the amount of alkaline phosphataseproduced is measured by a standard colourimetric assay. Dose responsecurves were constructed from which EC₅₀ values were estimated.

In this test the compound of formula (I) showed an EC₅₀ value of <1 nM.

The glucocorticoid receptor (GR) can function in at least two distinctmechanisms, by upregulating gene expression through the direct bindingof GR to specific sequences in gene promotors, and by downregulatinggene expression that is being driven by other transcription factors(such as NFκB or AP-1) through their direct interaction with GR.

In a variant of the above method, to monitor these functions, tworeporter plasmids have been generated and introduced separately intoA549 human lung epithelial cells by transfection. The first cell linecontains the firefly luciferase reporter gene under the control of asynthetic promoter that specifically responds to activation of thetranscription factor NFκB when stimulated with TNFα. The second cellline contains the renilla luciferase reporter gene under the control ofa synthetic promotor that comprises 3 copies of the consensusglucocorticoid response element, and which responds to directstimulation by glucocorticoids. Simultaneous measurement oftransactivation and transrepression was conducted by mixing the two celllines in a 1:1 ratio in 96 well plate (40,000 cells per well) andgrowing overnight at 37° C. Test compounds were dissolved in DMSO, andadded to the cells at a final DMSO concentration of 0.7%. Afterincubation for 1 h 0.5 ng/ml TNFα (R&D Systems) was added and after afurther 15 hours at 37° C., the levels of firefly and renilla luciferasewere measured using the Packard Firelite kit following themanufacturers' directions. Dose response curves were constructed fromwhich EC₅₀ values were determined. Transactivation (GR) Transrepression(NFκB) ED₅₀ (nM) ED₅₀ (nM) Compound of Formula (I) 0.06 0.20 Metabolite(X) >250 >1000 Fluticasone propionate 0.07 0.16

In Vivo Pharmacological Activity

Pharmacological activity in vivo was assessed in an ovalbumin sensitisedBrown Norway rat eosinophilia model. This model is designed to mimicallergen induced lung eosinophilia, a major component of lunginflammation in asthma.

For the experiments in this section, compound of formula (I) was used asunsolvated Form 1.

Compound of formula (I) produced dose dependant inhibition of lungeosinophilia in this model after dosing as an intra-tracheal (IT)suspension in saline 30 min prior to ovalbumin challenge. Significantinhibition is achieved after a single dose of 30 μg of compound offormula (I) and the response was significantly (p=0.016) greater thanthat seen with an equivalent dose of fluticasone propionate in the samestudy (69% inhibition with compound of formula (I) vs 41% inhibitionwith fluticasone propionate).

In a rat model of thymus involution 3 daily IT doses of 100 μg ofcompound (I) induced significantly smaller reductions in thymus weight(p=0.004) than an equivalent dose of fluticasone propionate in the samestudy (67% reduction of thymus weight with compound (I) vs 78% reductionwith fluticasone propionate).

Taken together these results indicate a superior therapeutic index forcompound (I) compared to fluticasone propionate.

In vitro Metabolism in Rat and Human Hepatocytes

Incubation of compound (I) with rat or human hepatocytes shows thecompound to be metabolised in an identical manner to fluticasonepropionate with the 17-β carboxylic add (X) being the only significantmetabolite produced. Investigation of the rate of appearance of thismetabolite on incubation of compound (I) with human hepatocytes (37° C.,10 μM drug concentration, hepatocytes from 3 subjects, 0.2 and 0.7million cells/mL) shows compound (I) to be metabolised ca. 5-fold morerapidly than fluticasone propionate:- 17-β acid metabolite productionSubject Cell density (pmol/h) number (million cells/mL) Compound (I)Fluticasone propionate 1 0.2 48.9 18.8 1 0.7 73.3 35.4 2 0.2 118 9.7 20.7 903 23.7 3 0.2 102 6.6 3 0.7 580 23.9

Median metabolite production 102-118 pmol/h for compound (I) and18.8-23.0 pmol/h for fluticasone propionate.

Pharmacokinetics after Intravenous (IV) and Oral Dosing in Rats

Compound (I) was dosed orally (0.1 mg/kg) and IV (0.1 mg/kg) to maleWistar Han rats and pharmacokinetic parameters determined. Compound (I)showed negligible oral bioavailability (0.90%) and plasma clearance of47.3 mL/min/kg, approaching liver blood flow (plasma clearance offluticasone propionate=45.2 mL/min/kg).

Pharmacokinetics after Intra-Tracheal Dry Powder Dosing in the Pig.

Anaesthetised pigs (2) were dosed intra-tracheally with, a homogenousmixture of compound (I) (1 mg) and fluticasone propionate (1 mg) as adry powder blend in lactose (10% w/w). Serial blood samples were takenfor up to 8h following dosing. Plasma levels of compound (I) andfluticasone propionate were determined following extraction and analysisusing LC-MS/MS methodology, the lower limits of quantitation of themethods were 10 and 20 pg/mL for compound (I) and fluticasone propionaterespectively. Using these methods compound (I) was quantifiable up to 2hours after dosing and fluficasone propionate was quantifiable up to 8hours after dosing. Maximum plasma concentrations were observed for bothcompounds within 15 min after dosing. Plasma half-life data obtainedfrom IV dosing (0.1 mg/kg) was used to calculate AUC (0-inf) values forcompound (I). This compensates for the plasma profile of Compound (I)only being defined up to 2 hours after an IT dose and removes any biasdue to limited data between compound (I) and fluticasone propionate.

C_(max) and AUC (0-info values show markedly reduced systemic exposureto compound (I) compared to fluticasone propionate:- Cmax AUC (0-inf)(pg/mL) (hr · pg/mL) Pig 1 Pig 2 Pig 1 Pig 2 Compound of Formula (I) 117 81 254 221 Fluticasone propionate 277 218 455 495

The pharmacokinetic parameters for both compound (I) and fluticasonepropionate were the same in the anaesthetised pig following intravenousadministration of a mixture of the two compounds at 0.1 mg/kg. Theclearance of these two glucocorticoids is similar is this experimentalpig model.

EXAMPLES Example 1 Nasal Formulation Containing6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicAcid S-fluoromethyl Ester

A formulation for intranasal delivery was prepared with ingredients asfollows: 6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-  0.05% w/w11β-hydroxy-16α-methyl-3-oxo- androsta-1,4-diene-17β-carbothioic acidS-fluoromethyl ester (prepared according to Intermediate 3, micronisedto MMD 3 μm) Polysorbate 80 0.025% w/w Avicel RC591  1.5% w/w Dextrose 5.0% w/w BKC 0.015% w/w EDTA 0.015% w/w water to 100%in a total amount suitable for 120 actuations and the formulation wasfilled into a bottle (plastic or glass) fitted with a suitablepre-compression pump and nasal actuator, adapted to dispense 50 or 100μl per actuation, preferably 50 μl

The formulation was prepared following the following protocol:

Part A

-   -   1. Dissolve dextrose in purified water    -   2. Dissolve EDTA in dextrose solution    -   3. Add Avicel RC591 while stirring    -   4. Allow suspension to hydrate        Part B (separately)    -   1. Dissolve polysorbate 80 in purified water at 50-60 ° C.    -   2. Prepare slurry of drug in Polysorbate 80 solution        Part C    -   1. Combine suspension of A4 with suspension of B2 and stir    -   2. Add solution of BKC in purifed water and stir    -   3. Adjust pH with 1N HCl    -   4. Add purified water to correct weight

Example 2 Nasal Formulation Containing6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17β-carbothioicAcid S-fluoromethyl Ester

A formulation for intranasal delivery was prepared with ingredients asfollows: 6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-  0.1% w/w11β-hydroxy-16α-methyl-3-oxo- androsta-1,4-diene-17β-carbothioic acidS-fluoromethyl ester (prepared according to Intermediate 3, micronisedto MMD 3 μm) Polysorbate 80 0.025% w/w Avicel RC591  1.5% w/w Dextrose 5.0% w/w BKC 0.015% w/w EDTA 0.015% w/w water to 100%in a total amount suitable for 120 actuations and the formulation wasfilled into a bottle (plastic or glass) fitted with a suitablepre-compression pump and nasal actuator, adapted to dispense 50 or 100μl per actuation, preferably 50 μl.

Example 3 Nasal Formulation Containing6α,9α-Difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxo-androsta-1,4-diene-17βcarbothioicAcid S-fluoromethyl Ester

A formulation for intranasal delivery was prepared with ingredients asfollows: 6α,9α-Difluoro-17α-1[(2-furanylcarbonyl)oxy]-  0.1% w/w11β-hydroxy-16α-methyl-3-oxo-androsta-1,4- diene-17β-carbothioic acidS-fluoromethyl ester (prepared according to intermediate 3, micronisedto MMD, 1.5 μm) Polysorbate 80 0.025% w/w Avicel RC591  1.5% w/wDextrose  5.0% w/w BKC 0.015% w/w EDTA 0.015% Water to 100%in a total amount suitable for 120 actuations and the formulation wasfilled into a bottle (plastic or glass) fitted with a suitablepre-compression pump and nasal actuator, adapted to dispense 50 or 100μl per actuation. Preferably 50 μl.

The formulation was prepared following the different protocol below:

Part A

-   -   5. Dissolve dextrose in purified water    -   6. Dissolve EDTA in dextrose solution    -   7. Add Avicel RC591 while stirring    -   8. Allow suspension to hydrate        Part B (separately)    -   3. Dissolve polysorbate 80 in purified water at 50-60° C.    -   4. Prepare slurry of drug in polysorbate 80 solution using a        suitable micronfluidizer, e.g. Microfluidics 110S, ensuring        suitable processing to attain target MMD of ,1.5 μm        Part C    -   5. Combine suspension of A4 with suspension of B2 and stir    -   6. Add solution of BKC in purified water and stir    -   7. Adjust pH with 1N HCl    -   8. Add purified water to correct weight.

Stability studies on Example 3 showed it to be stable up to 3 months at40° C. (measurements were not taken beyond this time). The particle sizeof the active material has also been observed not to changesignificantly.

The device of the invention will now be described, by way of exampleonly, with reference to the accompanying drawing in which:

FIG. 1 shows an exploded view of a nasal inhaler suitable for use inaccord with the invention;

With reference to FIG. 1, there is shown a nasal inhaler device 5comprising a body 6 a container 3 and a nasal pump 8. The device furthercomprises a protective end cap 7 having an inner surface 4 forengagement with the body 6 to protect the dispensing nozzle 11.

The body 6 is made from a plastic material and defines a housing 9 and adispensing nozzle 11. The housing 9 defines a cavity formed by a sidewall and a first end wall and a second end wall 14. The dispensingnozzle 11 is connected to and extends away from the second end wall 14and has an external tapering form.

The dispensing nozzle 11 has a longitudinally extending orifice definedby an outlet tube extending towards the cavity 10. An annular abutmentis formed within the orifice part way along the outlet tube. The annularabutment defines a small aperture through which fluid can flow in use.

The nasal pump 8 comprises a hollow casing 30 defining a reservoircontaining several doses of the fluid nasal formulation to be dispensedand a plunger slidably engaged within the hollow casing 30. One detailedexample of a suitable nasal pump Is described in U.S. Pat. No. 4,964,069incorporated herein by reference.

Throughout the specification and the claims which follow, unless thecontext requires otherwise, the word ‘comprise’, and variations such as‘comprises’ and ‘comprising’, will be understood to imply the inclusionof a stated integer or step or group of integers but not to theexclusion of any other integer or step or group of integers or steps.

The patents and patent applications described in this application areherein incorporated by reference.

1-4. (canceled)
 5. A pharmaceutical formulation comprising apharmaceutical formulation which comprises: an aqueous suspension ofparticulate compound of formula (I)

or a solvate thereof, and one or more preservatives, wherein thepreservative comprises disodium edatate. 6-8. (canceled)
 9. Apharmaceutical formulation according to claim 5 which further comprisesone or more wetting agents.
 10. A pharmaceutical formulation accordingto claim 9 wherein the wetting agent comprises polyoxyethylene (20)sorbitan monooleate.
 11. A pharmaceutical formulation according to claim9 wherein the wetting agent is present within the formulation in anamount of between 0.001 and 0.05% (w/w), based on the total weight ofthe formulation.
 12. A pharmaceutical formulation according to claim 5which further comprises: one or more isotonicity adjusting agents.
 13. Apharmaceutical formulation according to claim 12 wherein the isotonicityadjusting agent comprises dextrose.
 14. A pharmaceutical formulationaccording to claim 12 wherein the isotonicity adjusting agent is presentwithin the formulation in an amount of between 0.1 and 10% (w/w), basedon the total weight of the formulation.
 15. A pharmaceutical formulationaccording to claim 12 characterised in that it is isotonic with fluidsof the nasal cavity.
 16. A pharmaceutical formulation according to claim12 which is pH adjusted to between 5 and
 7. 17. A pharmaceuticalformulation according to claim 16 which is pH adjusted usinghydrochloric acid and/or sodium hydroxide
 18. A pharmaceuticalformulation according to claim 5 wherein the compound of formula (I) orsolvate thereof is present within the formulation in an amount between0.005% and 1% (w/w), based on the total weight of the formulation.
 19. Apharmaceutical formulation according to claim 5 wherein the particles ofthe compound of formula (I) have a mass mean diameter of between 0.5 and10 μm.
 20. A pharmaceutical formulation according to claim 19 whereinthe particles of the compound of formula (I) have a mass mean diameterof between 1 and 5 μm.
 21. A pharmaceutical formulation according toclaim 5 wherein the compound of formula (I) is in the form of unsolvatedForm 1 polymorph.
 22. A pharmaceutical formulation which comprises: anaqueous suspension of particulate compound of formula (I)

or a solvate thereof wherein the particle size of the compound offormula (I) has been reduced.
 23. A pharmaceutical composition asclaimed in claim 22 wherein the particle size has been reduced bymicronisation.
 24. A pharmaceutical composition as claimed in claim 22wherein the particle size has been reduced by microfluidisation.
 25. Apharmaceutical formulation as claimed in claim 22, wherein the compoundof formula (I) was substantially in the form of equant or substantiallyequant particles, prior to particle size reduction.
 26. A pharmaceuticalformulation as claimed in claim 22 wherein the compound of formula (I)was substantially in the form of tetragonal bipyramidal or substantiallytetragonal bipyramidal particles prior to particle size reduction.
 27. Apharmaceutical formulation as claimed in claim 23 wherein the compoundof formula (I) is substantially in the form of acicular particles.
 28. Apharmaceutical formulation according to claim 22 which comprises (i) oneor more suspending agents; (ii) one or more preservatives; (iii) one ormore wetting agents; and (iv) one or more isotonicity adjusting agents29. A pharmaceutical formulation according to claim 28 wherein thesuspending agent is microcrystalline cellulose and carboxymethylcellulose sodium, the preservative is EDTA and benzalkoniumchloride, the wetting agent is polyoxyethylene (20) sorbitan monooleateand the isotonicity adjusting agent is dextrose.
 30. A pharmaceuticalformulation according to claim 22 which comprises anothertherapeutically active agent.
 31. A pharmaceutical formulation accordingto claim 30, wherein the other therapeutically active agent is ananti-histamine or an anti-allergic.
 32. A container comprising apharmaceutical formulation which comprises: an aqueous suspension ofparticulate compound of formula (I)

or a solvate thereof wherein the particle size of the compound offormula (I) has been reduced.
 33. A device adapted for intranasaldelivery of a pharmaceutical formulation comprising a containeraccording to claim
 32. 34. A compound of formula (I)

substantially in the form of equant or substantially equant particles.35. A compound as claimed in claim 34 wherein the particles are in theform of tetragonal bipyramidal or substantially tetragonal bipyramidalparticles.
 36. A compound of formula (I)

substantially in the form of acicular particles.
 37. A method oftreatment of allergic rhinitis which comprises administering to apatient a pharmaceutically acceptable amount of a formulation accordingto claim
 22. 38. The method according to claim 37 wherein theadministration is once-per-day.
 39. A pharmaceutical formulation asclaimed in claim 22 for use in human or veterinary medicine in thetreatment of patients with an inflammatory and/or an allergic condition.40. A pharmaceutical formulation as claimed in claim 39 for use intreatment once-per-day.
 41. A process for the manufacture of amedicament for the treatment of patients with an inflammatory and/orallergic condition comprising preparing a pharmaceutical formulationaccording to claim
 22. 42. (canceled)
 43. A process for the preparationof a compound of formula (I) substantially in the form of equant orsubstantially equant particles which process comprises dissolving acompound of formula (I) in a suitable solvating solvent andcrystallising the compound.
 44. A process as claimed in claim 43 whereinthe solvating solvent is acetone, propan-2-ol, ethylacetate ortetrahydrofuran.
 45. A process as claimed in claim 44 wherein thesolvating agent is acetone pr propan-2-ol.
 46. A process as claimed inclaim 43 wherein the compound of formula (I) is substantially in theform of tetragonal bipyramidal particles.